RNA-Seq of AML12 cells
Total RNA was extracted using Trizol reagent following the manufacturers procedure. The total RNA quantity and purity were analyzed with a Bioanalyzer 2100 and an RNA 1000 Nano LabChip Kit (Agilent, CA, USA), with RIN number >7.0. Poly(A). RNA was purified from total RNA (5 g) using poly-T oligo-attached magnetic beads using two rounds of purification. Following purification, the mRNA was fragmented into small pieces using divalent cations under an elevated temperature. Then the cleaved RNA fragments were reverse-transcribed to create the final cDNA library in accordance with the protocol for the mRNA-Seq sample preparation kit (Illumina, San Diego, CA, USA). The paired-end sequencing was performed on an Illumina Novaseq 6000 (LC Sciences, USA), following the vendors recommended protocol. The sequencing work was commissioned by Lianchuan Biotechnology Co., LTD, Hangzhou, China.